Things we do ...
W e work primarily with mouse models where we can examine T cell behavior in vivo at a high resolution. We use transgenic and knockout mice of various flavors routinely - and are involved in generating some of our own. The mechanisms we study are (hopefully) relevant to a wide variety of patho-physiological contexts. So, depending on the specific experimental question, we have available in the lab, the following models ::
Autoimmunity :
- Arthritis
- Psoriasis / Dermatitis
- Hyperacute rejection
- Solid Graft rejection
Infectious disease:
- Leishmania
- Plasmodium
- Melanoma
The lab uses a wide variety of cellular and molecular techniques as required by particular experiments. On any given week we may be doing any or all of these ...
- Isolation of primary T, B and Dendritic cells, magnetic bead purifications, MACS etc.
- Flow Cytometry & Cell sorting
- Overexpression/Knockdown of genes in primary lymphocytes using Amaxa nucleofaction, Retroviral transduction or Electroporation
- Retrogenic mice - retroviral transduction of primary Bone Marrow stem cells followed by generation of chimeric mice (primarily to clone & express specific T cell receptors)
- ELISAs, Luminex, Bead array & other cytokine measurements
- TCR repertoire analysis, MHC tetramers, expression in insect cells
- Culture, growth and infection with parasites
- Cell culture and in vitro assays for T or DC function
- Standard molecular biology & biochemistry techniques.
R
eagents from the lab:
- Plasmids generated by us are available from the Addgene repository: https://www.addgene.org/Nevil_Singh/
- Contact the Lab directly for any mouse strains.